Action and mechanism of GATA2 overexpression regulating the transcriptome expression of chicken preadipocytes
Objective To clarify the impact of GATA2 overexpression on transcriptional expression in chicken preadipocytes.Methods Chicken preadipocytes ICP1 were cultivated,pCMV-myc-GATA2 or pCMV-myc plasmid was transfected,Western blot was used to verify the overexpression of GATA2 in cells,RNA-seq was used to study the changes in cell transcriptome expression caused by overexpression of GATA2,ChIP-seq was used to study the binding of GATA2 in the genome,Real time PCR was used to verify some differentially expressed genes,ChIP-PCR was used to verify the collection of GATA2 to TAF3 genes.Results Transfection of pCMV-myc-GATA2 allowed overexpression of GATA2 protein in ICP1 cells;overexpression of GATA2 resulted in up-regulation of 942 genes and down-regulation of 840 genes in ICP1 cells(P＜0.05),and enrichment analysis showed that differentially ex-pressed genes were associated with ribosome and mitochondrial structure and function(P＜0.01);GESA analysis showed that the expression of GATA2-overexpressing ICP1 cells showed up-regulation of RNA polymerase Ⅱ tran-scription initiation complex,ribosome biogenesis,propionate metabolism,and oxidative phosphorylation-related gene sets,and down-regulation of histone lysine N-methyltransferase,nuclear transcription repression complex formation,adipokines,and calcium ions signaling pathway-related gene sets.ChIP-seq in ICP1 cells identified 2833 GATA2-binding genomic peaks involving 2018 genes.motif analysis revealed GATA2-binding(T/A)GATA motifs;enrich-ment analysis showed that these genes are involved in embryonic development,signaling,cellular metabolism,and cell-cell interactions.Differently expressed genes and GATA2-binding genes were taken to intersect to obtain 105 genes,and enrichment analysis showed that these genes were associated with transcription,post-transcriptional regu-lation,cell-cell interactions,signaling,and cell cycle(P＜0.001).Conclusion GATA2 can at least bind to the genome of chicken preadipocytes to regulate their transcriptome expression patterns.
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